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. Author manuscript; available in PMC: 2022 Apr 1.
Published in final edited form as: Adv Ther (Weinh). 2021 Jan 20;4(4):2000244. doi: 10.1002/adtp.202000244

Figure 3. Cell uptake assessments reveal three promising dg-Bcan-targeting peptides (BTP).

Figure 3.

a) Table summarizing the properties of the seven peptides from the OBOC library screen. Each letter abbreviates a d-amino acid. MW = molecular weight. b) Chemical structure of the peptide labeled with fluorescein dye. c) Western blot of dg-Bcan and Bcan in human embryo kidney (HEK) cells, and Bcan-overexpressing HEK cells (HEK-Bcan). d) Flow cytometry analysis showing cell uptake of fluorescein-labeled peptides with or without blocking (50x excess native peptide) in HEK-Bcan cells (nevents = 20,000, two-way ANOVA). e) Cell uptake analysis of fluorescein-labeled peptides in BT188 (dg-Bcan-high) or BT145 (dg-Bcan-low) patient derived glioma cells (PDGCs) as measured by fluorescence microscopy. (nPDGCs = 10, two-way ANOVA). f) Chemical structure of the peptide labeled with Cy5.5 dye. g) Western blot analysis showing dg-Bcan expression in GBM-6 cells, astrocytes and neural progenitor cells (NPCs). h) Flow cytometry analysis showing GBM-6 cell uptake of the top 3 candidates, BTP-7-Cy5.5, BTP-8-Cy5.5 and BTP-9-Cy5.5 (each group normalized to a control scramble peptide (Scr-8-Cy5.5) (nevents = 20,000, two-way ANOVA) significance measured in comparison to Scr-8-Cy5.5). i) Flow cytometry analysis showing cell uptake of BTP-7-Cy5.5, BTP-8-Cy5.5 and BTP-9-Cy5.5 in GBM-6 cells and NPCs (nevents = 20,000 cells, two-way ANOVA).